Great human CCR5 staining antibody for flow cytometry

November 18, 2016

IMS
University of Toronto
PhD Candidate

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

anti-Human CCR5 (CD195) PE

Catalog Number:

359105

This antibody brightly stains human CCR5 on human lymphocytes such as CD4 T cells. CCR5 is a co-receptor for HIV infection, hence staining for CCR5 of lymphocytes are indicative of their susceptibility to HIV infection. This BioLegend antibody stains CCR5 on the surface and the positive population can be determined based off gating on controls such as isotype control or FMO. Shown is antibody staining on a T cell line (MOLT-4) that has been transfected with a CCR5 plasmid, maintained in antibiotic to ensure maximal, constitutive CCR5 expression.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human T cell line (MOLT-4) transfected with human CCR5 expression plasmid

Primary Incubation

5uL of antibody in 100uL staining buffer for 30 min at 4degC

Blocking Agent

2% FBS in PBS

Secondary Incubation

None

Tertiary Incubation

None

Detection

PE channel on any flow cytometer

Results Summary

MOLT-4 human T cells were transfected with a CCR5 expression and cultured in antibiotic (G418) to ensure constitutive expression of human CCR5 on the surface of these cells. Cells were either left unstained/FMO as a control or stained with 5uL in 100uL of BioLegend Cat#359105 for 30 mins in 2% FBS in PBS buffer at 4DegC. Cells were then fixed in 2% Formalin and analyzed on a BD FACSCalibur. Shown are the unstained cells in red in the histogram over layed with the stained sample in blue. The antibody efficiently detects the bulk population to be CCR5+ as expected.

DOI or PMID #

N/A

Additional Notes

None

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Summary

The Good

Ease of use, staining is bright

The Bad

CCR5 detection is based over background staining - this is conventional for any CCR5 detection on flow cytometry.

The Bottom Line

Great, efficient antibody. Is a staple in any HIV lab looking to study HIV acquisition of CD4 T cells and also their risk of susceptibility. Surface stain is bright and works at suggested dilution by company (5/100).

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